How to Choose the Right Inducible Gene Expression System for Mammalian Studies?

Inducible gene expression systems are favored over stable expression systems in a wide variety of basic and applied research areas, including functional genomics, gene therapy, tissue engineering, biopharmaceutical protein production and drug discovery. This is because they are mostly reversible and thus more flexible to use. Furthermore, compared to constitutive expression, they generally exhibit a higher efficiency and have fewer side effects, such as cell death and delayed growth or development. Empowered by decades of development of inducible gene expression systems, researchers can now efficiently activate or suppress any gene, temporarily and quantitively at will, depending on experimental requirements and designs. Here, we review a number of most commonly used mammalian inducible expression systems and provide basic standards and criteria for the selection of the most suitable one.

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Inducible gene expression systems and plant biotechnology

Biotechnology Advances ◽

10.1016/j.biotechadv.2009.05.006 ◽

2009 ◽

Vol 27 (6) ◽

pp. 733-743 ◽

Cited By ~ 61

Author(s):

Giandomenico Corrado ◽

Marianthi Karali

Keyword(s):

Gene Expression ◽

Plant Biotechnology ◽

Expression Systems ◽

Inducible Gene Expression ◽

Inducible Gene

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High efficiency inducible gene expression system based on activation of a chimeric transcription factor in transgenic pine

Plant Cell Reports ◽

10.1007/s00299-005-0009-1 ◽

2005 ◽

Vol 24 (10) ◽

pp. 619-628 ◽

Cited By ~ 5

Author(s):

Wei Tang ◽

Ronald J. Newton ◽

Thomas M. Charles

Keyword(s):

Gene Expression ◽

Transcription Factor ◽

High Efficiency ◽

Expression System ◽

Inducible Gene Expression ◽

Gene Expression System ◽

Chimeric Transcription Factor ◽

Inducible Gene

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Cumate-Inducible Gene Expression System for Sphingomonads and Other Alphaproteobacteria

Applied and Environmental Microbiology ◽

10.1128/aem.02296-13 ◽

2013 ◽

Vol 79 (21) ◽

pp. 6795-6802 ◽

Cited By ~ 40

Author(s):

Andreas Kaczmarczyk ◽

Julia A. Vorholt ◽

Anne Francez-Charlot

Keyword(s):

Gene Expression ◽

Caulobacter Crescentus ◽

Paracoccus Denitrificans ◽

Expression System ◽

Model Organisms ◽

Inducible Gene Expression ◽

Gene Expression System ◽

Content Type ◽

Wide Range ◽

Inducible Gene

ABSTRACTTunable promoters represent a pivotal genetic tool for a wide range of applications. Here we present such a system for sphingomonads, a phylogenetically diverse group of bacteria that have gained much interest for their potential in bioremediation and their use in industry and for which no dedicated inducible gene expression system has been described so far. A strong, constitutive synthetic promoter was first identified through a genetic screen and subsequently combined with the repressor and the operator sites of thePseudomonas putidaF1cym/cmtsystem. The resulting promoter, termed PQ5, responds rapidly to the inducer cumate and shows a maximal induction ratio of 2 to 3 orders of magnitude in the different sphingomonads tested. Moreover, it was also functional in otherAlphaproteobacteria, such as the model organismsCaulobacter crescentus,Paracoccus denitrificans, andMethylobacterium extorquens. In the noninduced state, expression from PQ5is low enough to allow gene depletion analysis, as demonstrated with the essential genephyPofSphingomonassp. strain Fr1. A set of PQ5-based plasmids has been constructed allowing fusions to affinity tags or fluorescent proteins.

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Implementation of the agmatine-controlled expression system for inducible gene expression in Lactococcus lactis

Microbial Cell Factories ◽

10.1186/s12934-015-0399-x ◽

2015 ◽

Vol 14 (1) ◽

Cited By ~ 12

Author(s):

Daniel M. Linares ◽

Patricia Alvarez-Sieiro ◽

Beatriz del Rio ◽

Victor Ladero ◽

Begoña Redruello ◽

...

Keyword(s):

Gene Expression ◽

Lactococcus Lactis ◽

Expression System ◽

Inducible Gene Expression ◽

Inducible Gene

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Development of an inducible gene expression system for primary murine keratinocytes

Journal of Dermatological Science ◽

10.1016/j.jdermsci.2007.09.003 ◽

2008 ◽

Vol 49 (1) ◽

pp. 73-84 ◽

Cited By ~ 6

Author(s):

Priyadharsini Nagarajan ◽

Satrajit Sinha

Keyword(s):

Gene Expression ◽

Expression System ◽

Inducible Gene Expression ◽

Gene Expression System ◽

Inducible Gene

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Heat-Inducible Gene Expression System by Applying Alternating Magnetic Field to Magnetic Nanoparticles

ACS Synthetic Biology ◽

10.1021/sb4000838 ◽

2013 ◽

Vol 3 (5) ◽

pp. 273-279 ◽

Cited By ~ 25

Author(s):

Masaki Yamaguchi ◽

Akira Ito ◽

Akihiko Ono ◽

Yoshinori Kawabe ◽

Masamichi Kamihira

Keyword(s):

Gene Expression ◽

Magnetic Field ◽

Magnetic Nanoparticles ◽

Expression System ◽

Alternating Magnetic Field ◽

Inducible Gene Expression ◽

Gene Expression System ◽

Inducible Gene

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A tunable anthranilate-inducible gene expression system for Pseudomonas species

Applied Microbiology and Biotechnology ◽

10.1007/s00253-020-11034-8 ◽

2020 ◽

Vol 105 (1) ◽

pp. 247-258

Author(s):

Lena Hoffmann ◽

Michael-Frederick Sugue ◽

Thomas Brüser

Keyword(s):

Gene Expression ◽

Expression System ◽

Ant System ◽

Plant Host ◽

Inducible Gene Expression ◽

Gene Expression System ◽

Expression Levels ◽

Key Points ◽

Wide Range ◽

Inducible Gene

Abstract Pseudomonads are among the most common bacteria in soils, limnic ecosystems, and human, animal, or plant host environments, including intensively studied species such as Pseudomonas aeruginosa, P. putida, or P. fluorescens. Various gene expression systems are established for some species, but there is still a need for a simple system that is suitable for a wide range of pseudomonads and that can be used for physiological applications, i.e., with a tuning capacity at lower expression levels. Here, we report the establishment of the anthranilate-dependent PantA promoter for tunable gene expression in pseudomonads. During studies on P. fluorescens, we constructed an anthranilate-inducible AntR/PantA-based expression system, named pUCP20-ANT, and used GFP as reporter to analyze gene expression. This system was compared with the rhamnose-inducible RhaSR/PrhaB-based expression system in an otherwise identical vector background. While the rhamnose-inducible system did not respond to lower inducer concentrations and always reached high levels over time when induced, expression levels of the pUCP20-ANT system could be adjusted to a range of distinct lower or higher levels by variation of anthranilate concentrations in the medium. Importantly, the anthranilate-inducible expression system worked also in strains of P. aeruginosa and P. putida and therefore will be most likely useful for physiological and biotechnological purposes in a wide range of pseudomonads. Key points • We established an anthranilate-inducible gene expression system for pseudomonads. • This system permits tuning of gene expression in a wide range of pseudomonads. • It will be very useful for physiological and biotechnological applications.

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